Effects of inhibition of Rho/ROCK pathway on proliferation and migration of airway smooth muscle cells and related mechanisms

CUI Yun-Fei; LU Qing-Hua; HUANG Xiao; LIN Wei-Nan; HUANG Ting; YANG Qin.

doi:10.7499/j.issn.1008-8830.2405119

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Chinese Journal of Contemporary Pediatrics ›› 2024, Vol. 26 ›› Issue (9) : 974-981. DOI: 10.7499/j.issn.1008-8830.2405119

EXPERIMENTAL RESEARCH

Effects of inhibition of Rho/ROCK pathway on proliferation and migration of airway smooth muscle cells and related mechanisms

CUI Yun-Fei, LU Qing-Hua, HUANG Xiao, LIN Wei-Nan, HUANG Ting, YANG Qin.

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Abstract

Objective To investigate the effects and molecular mechanisms of inhibition of the Ras homolog gene (Rho)/Rho-associated coiled-coil forming protein kinase (ROCK) pathway on the proliferation and migration of airway smooth muscle cells involving myocardin (MYOCD). Methods Human airway smooth muscle cells were infected with the adenoviral vector Ad-ZsGreen-shRNA-hROCK1 in vitro. The cells were randomly divided into four groups: ROCK1 gene silencing control (shNC) group, shNC + arachidonic acid (AA, Rho/ROCK pathway activator) group, ROCK1 gene silencing (shROCK1) group, and shROCK1 + AA group (n=3 each). Quantitative real-time polymerase chain reaction and Western blot were used to detect the expression levels of ROCK1 and MYOCD mRNA and protein. ELISA was employed to measure the levels of globular actin and filamentous actin, while immunofluorescent staining and scratch assays were utilized to assess cell proliferation and migration. Results Compared to the shNC + AA group, the shROCK1 + AA group exhibited decreased levels of ROCK1 and MYOCD mRNA and protein expression, reduced expression levels of globular actin and filamentous actin, and diminished cell proliferation and migration capabilities (P<0.05). Conclusions Inhibition of the Rho/ROCK pathway suppresses the proliferation and migration of airway smooth muscle cells, which may be associated with the downregulation of MYOCD.

Key words

Asthma / Rho/ROCK pathway / Myocardin / Proliferation / Airway smooth muscle cell

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CUI Yun-Fei, LU Qing-Hua, HUANG Xiao, LIN Wei-Nan, HUANG Ting, YANG Qin.. Effects of inhibition of Rho/ROCK pathway on proliferation and migration of airway smooth muscle cells and related mechanisms[J]. Chinese Journal of Contemporary Pediatrics. 2024, 26(9): 974-981 https://doi.org/10.7499/j.issn.1008-8830.2405119

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